Volume 65 (2015): Issue 2 (June 2015)

During the acute phase response, there is an increased production and release of certain proteins known as acute phase proteins (APPs) which can be produced by hepatocytes and peripheral tissues such as C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (Hp), alpha-1 acid glycoprotein (AGP). These proteins have been investigated as markers of various infectious diseases in small animals and the purpose of this review is to update the current knowledge about APPs in infectious diseases in dogs and cats.

Damage to the small intestine and impairment of the intestinal epithelium occur in various diseases, resulting in a need for new epithelium. Therefore, bioengineering of the small intestine is becoming an attractive field of research where all contributions are highly appreciated.

The purpose of this study was to determine the possibility of the multiplication of the enterocyte mass using the technique of serosal patch with the application of hematopoietic stem cells, as well as the assessment of the quality of newly formed mucosa.

Sixty Mill Hill hooded rats were divided in 4 groups, 15 animals each. In the control group animals, the patch was not created. In the other three groups, the animals were operated on and in each group 8 parietal and 7 visceral patches have been created. One of the groups with operated animals (Group NS) was not postoperatively treated. The second group of operated animals (Group G) was stimulated with granulocyte colony-stimulating factor (G-CSF). The third group of operated animals (Group GM) was stimulated with recombined humane granulocyte-macrophage colony-stimulating factor (rHuGM-CSF).

In the group of animals that were not stimulated, epithelium proliferated slowly. In the group of animals stimulated with G-CSF stimulants, the epithelium initially proliferated rapidly, but appeared atrophic after eight weeks. Stimulation by rHuGM-CSF led to faster epithelization, and epithelium showed signs of advancing proliferation after eight weeks.

We confirmed the possibility of enterocyte mass multiplication by using the serosal patch technique, as well as that stimulation with rHuGM-CSF is more effective than stimulation with G-CSF.

This study was aimed to determine the effect of the rearing system on the establishment and development of different functional groups of microorganisms in the rumen of kid goats. Fifty kids were fed on goat milk until weaning at 45 (TR, traditional rearing system, n = 25) and 30 days of age (alternative rearing system, AR, n = 25). In addition, only AR group was offered with rumen starter from birth. Both groups consumed alfalfa hay and ground corn between 30 and 90 days of age. Five kids from each group were slaughtered at 21, 30, 45, 70 and 90 days old. It was determined the total number of protozoa, anaerobic, amylolytic and cellulolytic bacteria present in the rumen. Kids of AR were lighter in weight than TR kids between 42 and 56 days old. In both rearing systems, anaerobic and amylolytic bacteria were found at 21 days of age, while cellulolytic and protozoa were not found until 45 days of age. Kids of AR had higher quantities of anaerobic and amylolytic bacteria until 30 and 45 days of age, respectively. These results demonstrate the rearing system does not affect the sequence and time in which the functional groups of microorganisms are established in the rumen. However, the alternative rearing system with early intake of solid food allowed the establishment of greater amount of bacteria and protozoa. Nevertheless, the effect of weaning on growth rate was more marked in kids from alternative rearing system, despite its greater microbiological rumen development.

The aim of this investigation was to determine the influence of oxidative stress upon general stress in dairy cows on parturition and whether the indicators of stress can be reduced by selenium (Se) supplementation.

A total of 36 animals were divided into 3 groups 21 days prepartum and treated with a single-term intramuscular supplement of sodium selenite: Control group - 0 mg; group Se10 - 10 mg; group Se20 - 20 mg.

Se supplementation significantly raised blood Se content and glutathione peroxidase (GPx) activity in groups Se10 and Se20, compared to Control, although there was no marked difference between supplemented groups. Plasma malondialdehyde (MDA) and cortisol concentrations were significantly reduced in supplemented groups Se10 and Se20, compared to Control. A negative correlation was detected between blood GPx activity and plasma MDA, while a positive correlation was determined between plasma MDA and cortisol concentrations.

These results indicate that prepartum Se supplementation can be utilized for a partial relief of stress in cows during labor by augmenting the antioxidative action of GPx.

Escherichia coli is an opportunistic pathogen affecting bovine mammary gland causing mainly transient infections; however, some recent reports indicated that some strains are able to adhere to and internalize into the epithelial cells, which can result in the persistence of the pathogen in the tissue and development of recurrent mastitis. The mechanism of adaptation of E. coli to the mammary gland relies on structures that are distinctive components of its extracellular matrix - curli fimbriae (proteinaceous component) and cellulose (polysaccharide). Expression of these components varies among the isolates. In this study, we investigated the capacity of expression of curli fimbriae and cellulose (via colony morphotype on Congo Red agar) and ability of biofilm formation (microtiter plate test) in 25 strains of E. coli isolated from milk of cows with clinical mastitis. Phylogenetic grouping of the isolates was performed using PCR method based on detection of chuA, yjaA and TspE4-C2 amplicons. Antimicrobial susceptibility was examined using standard disk diffusion test. Production of both extracellular matrix components was established in 56%, and expression of curli fimbriae in 64% E. coli isolates. All isolates that produced curli fimbriae, demonstrated this ability at a temperature of 37°C, indicating the potential role of these adhesive structures in the pathogenesis of mastitis. The results of phylogenetic typing confirmed that E. coli strains isolated from milk of cows with mastitis are typical commensals mainly belonging to phylogenetic groups A and B1. All curli and curli/cellulose producing isolates formed biofilm under in vitro conditions. The biofilm potentially plays an important role in the development of persistent infections as well as recurrent clinical symptoms after antibiotic therapy in spite of quite good in vitro antimicrobial susceptibility of the agent.

The aim of this work is the comparison of sodium, calcium, magnesium and phosphorus concentration in pre-ovulatory follicles and follicular cysts and their concentration in the serum of sows. The research was conducted on sows (Polish Large White x Polish Landrace crossbred). Sodium concentration in cystic fluid was higher than in the follicular fluid (P<0.05). Sodium concentration in the serum of cysts-bearing sows was higher than in non-cysts-bearing sows (P<0.01). Differences were also observed between cysts and serum of cysts-bearing sows (P<0.01), as well as follicles and serum of non-cysts-bearing sows (P<0.01). Calcium concentration in the serum of non-cystsbearing sows was higher than in the case of cysts-bearing sows (P<0.01). Differences were also between calcium concentration in the cysts and its concentration in the serum of cysts-bearing sows (P<0.05). Magnesium concentration in the serum of cysts-bearing sows was lower than in non-cysts-bearing sows (P<0.01). Differences in magnesium concentration were also between cysts and serum of cysts-bearing sows (P<0.01), as well as between follicles and serum of non-cysts-bearing sows (P<0.05). Phosphorus concentration in the cysts was lower than in the follicles (P<0.05). It was also lower in the serum of non-cysts-bearing sows than in the follicles (P<0.05). The concentration of sodium, magnesium and phosphorus in the serum was positively correlated with the concentration of the mentioned macro-elements in ovarian structures (r = 0.66, P<0.01; r = 0.49, and r = 0.48, P<0.05, respectively). A negative correlation was found between the concentration of serum calcium and the ovarian structures (r = -0.47, P<0.05).

The differences in the concentrations of macro-elements suggest that the ionic composition may be one of the factors associated with the processes occurring in the ovarian structures of pigs.

Mast cell neoplasia in dogs can occur in two different forms: common as cutaneous tumor, or less common as a systemic form of neoplastic mast cell proliferation - systemic mastocytosis. The aim of this study was to compare the histological and cytological features, KIT receptor expression and presence of c-KIT proto-oncogene mutations in neoplastic cells of dogs with canine cutaneous mast cell tumor (CMCT) and systemic mastocytosis. Microscopical examination of the cytological smears obtained from all selected dogs revealed that cellular specimens were constituted mostly of round cells with a central nuclei and fine to coarse purple cytoplasmic granules. Histopathological examination of skin samples of dogs with CMCT and a dog with systemic mastocytosis showed proliferation of the neoplastic mast cells in the superficial and/or deep dermis. Similar findings were observed in tissue samples derived from lymph nodes, spleen, liver, myocardium and kidneys of a dog with systemic mastocytosis. Three dogs with high grade CMCT as well as one dog with systemic mastocytosis showed cytoplasmic CD117 expression, while 3 dogs with low grade CMCT, had membranous expression of CD117. Based on our study, histological features and cytoplasmic CD117 expression in neoplastic cells of dogs with systemic mastocytosis are similar to those in dogs with high grade CMCTs. Nevertheless, mutations of c-KIT proto-oncogene were not found in tumor samples either from dogs with CMCT or dog with systemic mastocytosis.

Merle patterning in dogs, caused by the insertion of a short interspersed element (SINE) in the genetic structure of SILV gene, is characterized by patches of diluted pigment intermingled with normal melanin. Sequencing analyses of SINE element localized in the canine SILV gene discovered a variability of the poly (A)-tail length which is responsible for the different expression of merle pattern. The SINE element with the length of poly(A)-tail between 91 - 101 nucleotides is responsible for the merle phenotype with all characters of merle pattern. On the contrary the dogs which have SINE element with the shorter length of poly(A) tail between 54-65 nucleotides are referred as cryptic merles without expresion of Merle pattern. The aim of this study was to improve molecular genetics method for the detection of cryptic allele for merle patterning in dogs. A total of 40 dogs of four breeds - Border collie, Shetland sheepdog, Australian Shepherd dog, and Chihuahua were used in this study. Canine genomic DNA was isolated from samples of whole blood and buccal cells by commercial column kit. Detection of merle (M), cryptic merle (Mc) and non-merle (m) alleles was done using M13-tailed primer protocol and two different allele-sizing methods for the verification of the electrophoresis result. In the analyzed population of dogs were detected 20 dogs with non-merle genotype mm, 17 dogs with merle genotype Mm, 2 dogs with double merle genotype MM and one dog with merle phenotype but with the presence of cryptic merle allele Mc with the consequential genotype MMc.

The purpose of this work was to investigate the influence of hypothyroidism on spleen tissue morphology and immune cell density in fourteen-day-old juvenile rats. Hypothyroidism in pups (n=10) was induced by administration of propylthiouracil (PTU) in drinking water (1.5 mg/L) to their mothers during pregnancy and period of lactation. Fourteen-day-old pups were sacrificed and the thyroid-stimulating hormone (TSH) serum concentration and thyroid activation index (Ia) were determined. Increased serum level of TSH and increased Ia showed that pups from PTU treated mothers were hypothyroid. White and red spleen pulp, marginal zone and connective tissue volume density has been assessed by using the stereological method. Using immunohistochemistry, the present CD3+ T lymphocytes, CD45RA+ B lymphocytes and CD68+ macrophages were quantified. A significant reduction of volume density of the periarteriolar lymphocyte sheath (VvPALS) and lymphatic follicles (Vvf) due to depletion of T and B lymphocytes respectively, was observed in the spleens of hypothyroid pups compared to controls. The volume density of the red pulp (Vvrp), marginal zone (Vvmz) and connective tissue (Vvct) was increased, as well as the number of CD68+ macrophages in the spleens of hypothyroid pups compared to controls. These results indicate that thyroid hormones might be important for normal development of both, specific and innate immune cells in the spleen during prenatal and early postnatal period.

The objective of this study was to describe the physiological changes in the concentrations of acute phase proteins (APPs) in calves during the first month of life, including pre-, postcolostral and milk feeding period. Seven clinically healthy calves were used in this study. Calves received colostrum and whole milk from their dams. The first blood sampling was performed before colostrum intake (day 0) and then at 1, 2, 7, 14 and 30 days of age. Blood serum was analyzed for the concentrations of haptoglobin (Hp), serum amyloid A (SAA), α1-acid glycoprotein (AGP), ceruloplasmin (Cp), and albumin (Alb). The results showed significant changes in the serum concentrations of Hp, SAA and Cp (P<0.001, P<0.01, P<0.01). Their lowest concentrations were found after birth, and a gradual increase was observed after colostrum intake until day 7 of life. Another trend was observed in the concentrations of albumin with a more marked decrease of values 1 day after colostrum intake and subsequent significant increase of values until the end of the first month of age (P<0.001). Sampling time had no significant effect on the concentrations of AGP. The values observed at birth and on day 1 of life were relatively stable. The concentrations of AGP increased slightly from day 2 until the end of the first month of age. These results suggest that the concentrations of APPs in the neonatal period are influenced by colostrum intake and age. This should be taken into consideration for the precise interpretation of these analytes in young animals.

The research was performed to determine the nutritive value of raw earthworms (Lumbricus rubellus) and dried earthworm powder, or earthworm meal (EM). In addition, the effects of a diet in which fish meal was substituted with EM or fresh earthworms on the health and productive performance of broilers were monitored. The experiment, which lasted 42 days, was conducted on one hundred Hybro G broilers divided into four equal groups. The control group was fed a standard feed, whereas, the first and the second one were given diets in which 50% or 100% of fish meal had been substituted with EM. The third experimental group received no fish meal but was given fresh earthworms ad libitum. The results of chemical analyses showed that earthworm meal contained 41.42% proteins (in dry matter) and satisfactory amounts of amino acids. Microbiological examination and tests for heavy metals suggest that earthworm meal and fresh earthworms did not contain heavy metals and harmful bacteria. Therefore, these feeds may be considered suitable for chickens. In the experiment as a whole no statistically significant difference (p>0.05) was observed in the productive performance between the experimental groups of broiler chickens. These results suggest that dehydrated earthworm meal can be an adequate substitute for fish meal in the broilers’ feed.

IFN-τ is a type I interferon, and it is known to be non-virus inducible in ruminants. IFN-τ reduced syncytium formation by PBMC obtained from BLV infected cattle in vitro. In order to clarify the effects of IFN-τ on cellular immune function in Japanese Black (JB) cattle with bovine leukemia virus (BLV) infection, immune related factors of peripheral blood mononuclear cells (PBMC) were analyzed using IFN-τ as a stimulator. Thirty-two JB cattle were used in this investigation, and these cattle were divided into three groups: cattle with enzootic bovine leucosis (EBL) (EBL Group, N=7), clinically healthy cattle with BLV infection (Carrier Group, N=13), and clinically healthy cattle without BLV infection (non-Carrier Group, N=12). A number of mRNA expressions of interleukin-12 and interferon (IFN)-as immune cell activating cytokines, perforin and granulysin as cytotoxic factors, and myxovirus resistance protein (MX)-1 and MX-2 as anti-virus factors of PBMC were analyzed after culturing cells with phytohemagglutinin (PHA) or IFN-τ. The basal mRNA levels of perforin and granulysin in the Carrier Group were significantly higher than those in the non-Carrier Group. Also, significantly higher basal mRNA levels of MX-1 and MX-2 in the EBL Group were detected compared with the non-Carrier Group. The mRNA expressions of perforin and granulysin in PBMC stimulated with PHA were higher in the Carrier Group than those in the non-Carrier Group. There were significantly higher mRNA levels of MX-1 and MX-2 in PBMC stimulated with IFN-τ in the EBL Group compared with those in the non-Carrier Group. These results suggest an enhanced sensitivity of anti-virus reaction in PBMC by IFN-τ treatment in JB cattle with EBL.